Preparation and processes for production thereof

Abstract

A meat protein hydrolysate derived from animal meat, wherein the protein hydrolysate is highly soluble in aqueous solution, exhibits low bitterness and is suitable for human consumption. The meat protein hydrolysate may be derived from animal meat, in a process that comprises: a) adding water or aqueous solution to ground or minced meat to produce an aqueous meat slurry; b) heating the aqueous meat slurry to from about 60 °C to about 75 °C for from about 10 mins to about 90 mins to produce a heated aqueous meat slurry; c) adjusting temperature of the heated aqueous meat slurry to protease functional temperature and adding sufficient protease to effect a first hydrolysis step, with stirring, over a period of from about 1 hr to about 3 hrs; d) adding sufficient peptidase to a product of the first hydrolysis step to effect a second hydrolysis step, with stirring, over a period of from about 30 mins to about 2 hrs; e) treating a product of the second hydrolysis step to deactivate the protease and / or peptidase to produce a meat hydrolysate mixture; f) exposing the meat hydrolysate mixture to one or more separation and / or purification steps to substantially remove solids and fats and produce meat protein hydrolysate.

Description

Preparation and processes for production thereofFIELD OF THE INVENTION

[0001] The present invention relates to the field of soluble meat hydrolysates and in particular, but not exclusively, to soluble meat hydrolysates that have improved sensory attributes. The invention also relates to processes for production of soluble meat hydrolysates and to products incorporating such hydrolysates. However, it will be appreciated that the invention is not limited to these particular fields of use.BACKGROUND OF THE INVENTION

[0002] The following discussion of the prior art is provided to place the invention in an appropriate technical context and enable the advantages of it to be more fully understood. It should be appreciated, however, that any discussion of the prior art throughout the specification should not be considered as an express or implied admission that such prior art is widely known or forms part of the common general knowledge in the field.

[0003] Hydrolysis of food proteins results in the formation of amino acids and peptides of a range of molecular weights, which are referred to as protein hydrolysates. Such materials are routinely utilised in the production of foods, beverages and supplements. For example, protein supplement powders, protein drinks, protein bars, soups and sauces, pre-prepared meals and weight gain / weight loss supplements and / or foods routinely include peptide and amino acid material derived from hydrolysis of food proteins.

[0004] Unfortunately, protein hydrolysates are often found to elicit bitter taste in the products in which they are incorporated. Depending upon the food protein from which the hydrolysate is derived, the hydrolysates and the products in which they are incorporated may include known human food allergens. The presence of such allergens may give rise to product safety issues and / or limit the commercial potential of the products including such material. This is especially likely when the protein hydrolysates are derived from milk, nuts, plants, eggs or shellfish, for example.

[0005] As outlined in References 1-3, bitterness is thought to be caused by the production of small peptides (generally MW 180 to 1000 Da or 2 to 10 amino acids) containing a higher number of hydrophobic amino acid residues, which are generally buried deep inside the folded tertiary structure of proteins and released upon hydrolysis of peptide bonds. In addition, hydrolysates derived from specific food protein utilising different proteases may vary in the level of bitterness due to differences in specificity of the proteases used in production.

[0006] A conventional process for production of enzyme hydrolysates of food protein may include steps such as:Grinding and dispersion of food proteinIEnzyme hydrolysis (at controlled pH, temperature and time)IEnzyme inactivationISeparation (e.g. centrifugation, decanting, filtration)ICollection of filtrate / supernatantIConcentration / drying

[0007] For example, W02009 / 077457 discloses a process for producing a partial hydrolysate of cereal protein with a degree of hydrolysis of from 9- 18%, intended to prevent allergic reactions in young mammals who consume the hydrolysate. This document discloses a pre-treatment heating step conducted at 60°C to 65°C for 10 minutes or at 40°C - 90°C for 1 to 8 hours. This document also discloses sequential treatment with different proteases, with an enzyme deactivation heating step conducted between. W02009 / 077457 does not disclose the problem of food protein hydrolysate bitterness or of means to improve the sensory attributes of such hydrolysates.

[0008] US5356637 discloses a method of preparing di- and tri-peptide rich hydrolysate, which are said to be important for intestinal absorption of protein derived material. The process involves pre-treating a protein mixture, which may be from slaughter blood, milk, fish isolate, soybean and lucerne and egg white, with heat before subsequent hydrolysis and then separation steps. An initial heat treatment is conducted at between 70-100°C for 30 to 45 mins and the process involves a single enzyme hydrolysis step. US5356637 does not disclose or address the problems of food protein hydrolysate bitterness or allergenicity.

[0009] CN 105248836 relates to production of a hypoallergenic whey protein powder produced in a two-step enzymatic hydrolysis process to produce ingredients for hypoallergenic infant milk powder. The process involves pre-treatment heating 80°C for 10 mins and two separate enzyme hydrolysis steps performed with different enzymes, each with an enzymedeactivation step (treatment at 75°C for 30 mins) conducted afterwards. CN 105248836 does not disclose or address the problem of food protein hydrolysate bitterness.

[0010] Preferably the present invention overcomes or to some extent ameliorates one or more of the disadvantages of the prior art, or at least provides a useful alternative.

[0011] A preferred embodiment of the present invention may provide a soluble meat protein hydrolysate that exhibits reduced bitterness compared to known food protein hydrolysate products. In preferred embodiments, the meat protein hydrolysates of the invention are also substantially devoid of egg, milk, nut, grain and legume derived human allergens.SUMMARY OF THE INVENTION

[0012] According to one aspect, the present invention provides a meat protein hydrolysate derived from animal meat, wherein the protein hydrolysate is highly soluble in aqueous solution, exhibits low bitterness and is suitable for human consumption.

[0013] Preferably, the meat protein hydrolysate has a degree of hydrolysis (DH) from about 10% to about 20% and preferably the meat protein hydrolysate has total soluble solids (TSS) from about 2% to about 6% w / w, in solution. The meat protein hydrolysate preferably also has a bitterness score of about 3 or less and preferably peptides within the hydrolysate have molecular weight of about 10kDa or less.

[0014] In one aspect the meat protein hydrolysate is in dry powder form. Preferably the hydrolysate has protein content of about 80% (w / w) or higher and fat content of about 1% (w / w) or lower and preferably the meat protein hydrolysate exhibits solubility of 95 % w / w or greater in aqueous solution at pH of from about 2 to about 8.

[0015] In another aspect of the invention the meat protein hydrolysate is in aqueous solution form.

[0016] For example, the meat protein hydrolysate may be produced from low-grade meat cuts and / or meat by- and co-products and the animal meat may for example be bovine, ovine, porcine, poultry, caprine, cameline, macropod or piscine.

[0017] In one aspect of the invention the meat protein hydrolysate is substantially devoid of egg, milk, nut, grain and legume derived human allergens.

[0018] In another aspect, the meat protein hydrolysate comprises one or more carriers and / or additives.

[0019] In a further aspect of the present invention there is provided a process for production of a meat protein hydrolysate derived from animal meat, wherein the protein hydrolysate is highly soluble in aqueous solution, exhibits low bitterness and is suitable for human consumption, wherein the process comprises:a) adding water or aqueous solution to ground or minced meat to produce an aqueous meat slurry;b) heating the aqueous meat slurry to from about 60 °C to about 75 °C for from about 10 mins to about 90 mins to produce a heated aqueous meat slurry;c) adjusting pH and temperature of the heated aqueous meat slurry to protease functional temperature and pH and adding sufficient protease to effect a first hydrolysis step, with stirring, over a period of from about 1 hr to about 3 hrs;d) exposing a product of the first hydrolysis step to sufficient peptidase to effect a second hydrolysis step, with stirring, over a period of from about 30 mins to about 2 hrs;e) treating a product of the second hydrolysis step to deactivate the protease and / or peptidase to produce a meat hydrolysate mixture;f) exposing the meat hydrolysate mixture to one or more separation and / or purification steps to substantially remove solids and fats and produce meat protein hydrolysate.

[0020] In one aspect of the invention steps c) and d) are conducted simultaneously and in another aspect steps c) and d) are conducted separately.

[0021] For example, the protease may comprise a neutral endoproteinase and the peptidase may comprise an exopeptidase.

[0022] In one aspect, in step b) the aqueous meat slurry is heated to about 70 °C for about 30 mins.

[0023] In another aspect of the invention the deactivation of the protease and / or peptidase in step e) is achieved by heating the product of the second hydrolysis step to from about 80 °C to about 90 °C for from about 10 mins to about 20 mins.

[0024] In further aspects of the invention the one or more separation and / or purification steps comprise:i. filtration;ii. clarification;iii. separation;iv. activated carbon treatment;v. ultrafiltration.

[0025] In one aspect of the invention, the one or more separation and / or purification steps further comprises nanofiltration.

[0026] In one aspect of the invention the meat protein hydrolysate produced by the one or more separation and / or purification steps is pasteurized and in a further aspect the pasteurized meat protein hydrolysate is concentrated to produce a concentrated meat protein hydrolysate solution.

[0027] In a further aspect of the invention the concentrated meat protein hydrolysate solution is spray dried to produce a meat protein hydrolysate powder.

[0028] In a further aspect of the present invention there is provided a meat protein hydrolysate produced by the process outlined above.

[0029] In another aspect of the invention there is provided a food, beverage or supplement product for human consumption comprising the meat protein hydrolysate. In one aspect the product is a protein drink; a protein supplement powder; a protein bar; a soup or sauce; a preprepared meal or weight gain or weight loss supplement / food.DEFINITIONS

[0030] In describing and claiming the present invention, the following terminology will be used in accordance with the definitions set out below. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments of the invention only and is not intended to be limiting.

[0031] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one having ordinary skill in the art to which the invention pertains.

[0032] Unless the context clearly requires otherwise, throughout the description and the claims, the terms “comprise”, “'comprising”, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”. For example, a composition, mixture, process or method that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, mixture, process or method.

[0033] The transitional phrase "consisting of’ excludes any element, step, or ingredient not specified. If in the claim, such would close the claim to the inclusion of materials other than those recited except for impurities ordinarily associated therewith. When the phrase "consisting of' appears in a clause of the body of a claim, rather than immediately following the preamble, it limits only the element set forth in that clause; other elements are not excluded from the claim as a whole.

[0034] The transitional phrase "consisting essentially of" is used to define a composition, process or method that includes materials, steps, features, components, or elements, in addition to those literally disclosed, provided that these additional materials, steps, features, components, or elements do not materially affect the basic and novel characteristic(s) of the claimed invention. The term "consisting essentially of' occupies a middle ground between "comprising" and "consisting of".

[0035] Where the applicant has defined an invention or a portion thereof with an open-ended term such as "comprising", it should be readily understood that (unless otherwise stated) the description should be interpreted to also describe such an invention using the terms "consisting essentially of" or "consisting of." In other words, with respect to the terms “comprising”, “consisting of”, and “consisting essentially of’, where one of these three terms is used herein, the presently disclosed and claimed subject matter may include the use of either of the other two terms. Thus, in some embodiments not otherwise explicitly recited, any instance of “comprising” may be replaced by “consisting of” or, alternatively, by “consisting essentially of”.

[0036] While reference may be made in this disclosure to the invention comprising a combination of a plurality of elements, it is also understood that this invention is regarded to comprise combinations which omit or exclude one or more of such elements, even if this omission or exclusion of an element or elements is not expressly stated herein, unless it is expressly stated herein that an element is essential to the applicant's combination and cannot be omitted. It is further understood that the related prior art may include elements from which this invention may be distinguished by negative claim limitations, even without any express statement of such negative limitations herein. It is to be understood, between the positive statements of applicant's invention expressly stated herein, and the prior art and knowledge of the prior art by those of ordinary skill which is incorporated herein even if not expressly reproduced here for reasons of economy, that any and all such negative claim limitations supported by the prior art are also considered to be within the scope of this disclosure and its associated claims, even absent any express statement herein about any particular negative claim limitations.

[0037] As used herein, with reference to numbers in a range of numerals, the terms "about," "approximately" and "substantially" are understood to refer to the range of -10% to +10% of thereferenced number, preferably -5% to +5% of the referenced number, more preferably -1 % to + 1 % of the referenced number, most preferably -0 .1 % to +0 .1 % of the referenced number. Moreover, with reference to numerical ranges, these terms should be construed as providing support for a claim directed to any number or subset of numbers in that range. For example, a disclosure of from 1 to 10 should be construed as supporting a range of from 1 to 8, from 3 to 7, from 1 to 9, from 3.6 to 4.6, from 3.5 to 9.9, from 8 to 10, and so forth.

[0038] The terms “preferred” and “preferably” refer to embodiments of the invention that may afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, and is not intended to exclude other embodiments from the scope of the invention.

[0039] The complete disclosures of the patents, patent documents and publications cited herein are incorporated by reference in their entirety as if each were individually incorporated.

[0040] Unless expressly stated to the contrary, "or" refers to an inclusive or and not to an exclusive or. For example, a condition A or B is satisfied by any one of the following: A is true (or present) and B is false (or not present), A is false (or not present) and B is true (or present), and both A and B are true (or present).

[0041] The term "and / or" used in the context of "X and / or Y" should be interpreted as "X," or "Y," or "X and Y." Similarly, "at least one of X or Y" should be interpreted as "X," or "Y," or "both X and Y."

[0042] The indefinite articles "a" and "an" preceding an element or component of the invention are intended to be non-restrictive regarding the number of instances (i.e., occurrences) of the element or component. Therefore "a" or "an" should be read to include one or at least one, and the singular word form of the element or component also includes the plural unless the number is obviously meant to be singular.

[0043] As used herein, wt.% refers to the weight of a particular component relative to total weight of the referenced composition or relative to a specified component.

[0044] It will be understood that use of the term “from” herein when referring to a range of numerical values encompasses the numerical values at each endpoint of the range. For example, a temperature of from 80 °C to 150 °C is inclusive of a temperature of 80 °C and a temperature of 150 °C.

[0045] Various features of the embodiments of the invention disclosed herein are, for brevity, described in the context of a single embodiment, but may also be provided separately or in anysuitable sub-combination. All combinations of the embodiments are specifically embraced by the illustrative embodiments disclosed herein just as if each and every combination was individually and explicitly disclosed. In addition, all sub-combinations listed in the embodiments describing such variables are also specifically embraced by the present compositions and are disclosed herein just as if each and every such sub-combination was individually and explicitly disclosed herein.BRIEF DESCRIPTION OF THE DRAWINGS

[0046] The aspects described above, as well as other apparent aspects, advantages, and objectives of the present invention are apparent from the detailed description below in combination with the drawing, in which:

[0047] Fig. 1 shows a calibration curve for the size exclusion chromatography (SEC) column used, which is an Agilent AdvanceBio SEC (7.8 x 300 mm, 2.7 pm).

[0048] Fig. 2 shows the size exclusion chromatography analysis performed on the meat protein hydrolysate produced according to the laboratory scale general process of Example 1.DETAILED DESCRIPTION

[0049] The skilled addressee will understand that the invention comprises the embodiments and features disclosed herein as well as all combinations and / or permutations of the disclosed embodiments and features.

[0050] In a broad aspect the present invention relates to a meat protein hydrolysate derived from animal meat, wherein the protein hydrolysate is highly soluble in aqueous solution, exhibits low bitterness and is suitable for human consumption. By reference to “meat protein hydrolysate” it is intended to convey that animal protein has been exposed to hydrolysis wherein peptide bonds between amino acids within the protein are cleaved with enzymes to produce shorter peptides and / or amino acids.

[0051] The protein source for the hydrolysates of the invention is animal meat. For example, the animal meat may be derived from bovine (e.g. cattle, oxen, bison and buffalo) ovine, porcine, poultry (including farmed avians such as chicken, duck, turkey, quail, pheasant and goose, for example), caprine, cameline, macropod (such as kangaroo and wallaby) or piscine animals. In preferred aspects of the invention the meat is derived from cattle, sheep, chicken, pig, goat and / or kangaroo.

[0052] The meat derived from such animals may for example be derived from muscle, connective tissue, skin, ligaments, tendons, internal organs such as heart, liver, lungs, spleen. Inmany instances the meat source of protein utilised in the invention will be derived from low grade meat cuts (that is low value muscle meat cuts that are less desirable for human consumption than prime cuts and which may be considered unduly fatty, tough or less tasty than prime cuts) and / or meat co- or by-products (waste) (for example, comprising offal, off-cuts from butchering of prime cuts and material that would otherwise be discarded or transformed into animal feed or fertiliser). In preferred aspects of the invention the meat from which the hydrolysate is derived is devoid (or at least substantially devoid such that no detectable levels are present) of material derived from eggs, animal milk, nuts, grains and legumes. Preferably also, the meat is not processed on equipment that has processed such substances, to substantially eliminate the possibility of known human allergens from these sources being present in the protein hydrolysates of the invention.

[0053] The meat protein hydrolysates of the present invention are highly soluble in aqueous solution, which will suit the hydrolysates to being incorporated into a wide variety of food, beverage and supplement products. In such products it is generally desirable for the hydrolysates to boost peptide and amino acid levels, but to do so in a manner that does not markedly affect the processing, taste, sensory perception or colour of the product. By being highly soluble the peptides and amino acids within the hydrolysates will also be in a form that is readily absorbed by the consumer.

[0054] For example, being highly soluble in aqueous medium may be quantified as exhibiting solubility of 95 wt.% or greater in aqueous solution at pH of from about 2 to about 8, and preferably having solubility of at least 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.2%, 98.4%, 98.6%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 99.95%. For example, the solubility of the meat protein hydrolysates of the invention can be determined by the method of Anema etal (2006) (Ref 6). In this method solutions of 5% w / w freeze dried hydrolysate powder are prepared in distilled water and mixed at room temperature for 30 minutes using a rotary mixer followed by centrifugation at 700 g for 10 minutes. The supernatant is dried at 105°C for 24 h and solubility is calculated as the percentage of the dried weight of the supernatant to the initial weight of dried hydrolysate.

[0055] A key advantage associated with the meat protein hydrolysates of the present invention is that they exhibit low bitterness, which is desirable so that the hydrolysates do not adversely affect the sensory characteristics of food, beverage or supplement products within which the hydrolysates are incorporated. Ultimately, bitterness is a characteristic that can be determined though consumer tasting. However, the degree of hydrolysis (DH %) and the total soluble solids measurement (TSS %) both provide useful proxies for bitterness.

[0056] DH % can be assayed using the method of Nielsen et al (2001) (Reference 4) and the TSS % can be estimated by determining refractive index and comparing to a standard curvegenerated from hydrolysates with known total soluble solids. Hydrolysates with low or high DH % values are generally considered less bitter. By comparison with tasting panel results, the present inventors have shown that a DH % of from about 8 % to about 20 %, such as from about 9 % to about 18%, from about 10 % to about 17 %, from about 10 % to about 16 % or preferably from about 10 % to about 15 % is generally favourable from a bitterness perspective, whereas DH % values of from about 20 % to about 25 % are often associated with unacceptably high bitterness.

[0057] Total soluble solids (TSS %) can be estimated by measuring refractive index using a refractometer and deriving TSS % from a standard curve generated from hydrolysates with known total soluble solids. The TSS % provides an estimate of expected yield once the hydrolysate has been dried and can also be used to estimate the progress of hydrolysis. The hydrolysate includes peptides and amino acids generated during hydrolysis, as well as other soluble compounds, such as salts. Higher TSS results are indicative of a high yield of soluble hydrolysate compounds from the starting substrate but are dependent on the weight of substrate and hydrolysis water used. While higher yields may be favourable economically, higher TSS values can lead to more bitter end products. In hydrolysates prepared from 20 and 40 wt.% meat slurries the inventors have shown from the sensory trials that TSS values of up to about 6, such as from about 2% to about 6%, from about 2.5% to about 5% or preferably from about 2.5 % to about 3.5 %, are generally the most acceptable, particularly from a bitterness perspective.

[0058] Bitterness can also be assessed by sensory analysis with a panel of tasters, an approach to which is outlined in detail in Example 1. The sensory analysis reported in Example 1 utilised a panel of five consumer tasters. Panelists were asked to rate their perceived levels of beefy flavour, saltiness, and bitterness intensity from 0 to 10 and to note other flavours that were present and overall comments. In the experience of the inventors from the sensory trials conducted, hydrolysates with bitterness ratings of 4 or lower are generally acceptable and those with bitterness ratings of 3 or lower are considered the most acceptable. Bitterness ratings above 5 are generally undesirable. While the sensory panel approach to bitterness assessment relies on the subjective assessment of tasters, the reliability of this approach to bitterness determination can be improved by increasing the number of panel members, for example to 10, 15, 20, 25, 30, 40 or even 50 or more panelists.

[0059] Without wishing to be bound by theory, the present inventors understand that aspects of the process they have developed for production of the meat protein hydrolysates contribute significantly to the removal of bitter proteins I peptides. In particular, the pretreatment heating of the meat slurry to from about 60 °C to about 75 °C for from about 10 mins to about 90 mins is believed to be significant in removal of bitter proteins I peptides. The inventors consider that the thermal pretreatment step results not only in pasteurization of the starting protein material but also in denaturation of meat proteins leading to opening of folded / globular (secondary and tertiary)protein structure to rapidly yield more extensive protein hydrolysis. However, the temperature and time of heat exposure is considered insufficient to initiate heat-induced protein hydrolysis.

[0060] Another processing aspect that the inventors consider likely to contribute to removal of bitter proteins I peptides, especially when combined with adoption of the pretreatment heating, is sequential (rather than simultaneous) conduct of protease and peptidase treatment steps. Again, without wishing to be bound by theory, the inventors consider that adopting sequential hydrolysis of protein (by broad specificity protease and then peptidase preparations) permits production of a large pool of peptides from protein by broad specificity protease of varying molecular weights and hydrophilicity / hydrophobicity. These peptides are then further modified by exposure to peptidase. The inventors note that simultaneous addition of protease and peptidase preparations can potentially lead to hydrolysis / inactivation of peptidases, which are also proteinaceous in nature, by the protease preparation, which may have negative implications for the debittering activity of the peptidase. In some embodiments, a step of thermal inactivation of protease may be conducted before the addition of peptidase for the second hydrolysis step.

[0061] Preferably the meat protein hydrolysates of the invention comprise peptides substantially having molecular weight of about 10kDa or less. For example, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or about 99.3% of peptides within the hydrolydates may have molecular weight of 10 kDa or less.

[0062] In one aspect of the invention the meat hydrolysates of the invention are provided in the form of a dry powder, which is a form that is convenient for packaging and transportation and is suitable for incorporation into a variety of products. Dry powder hydrolysates can be produced by concentration and drying of aqueous liquid containing hydolysates using standard techniques as further discussed below.

[0063] In some embodiments the dry powder hydrolysates may be combined with one or more carriers or other additives. Examples of carriers and additives that may be combined with the hydrolysates of the invention include, but are not limited to, bulking agents, colourants, flavouring agents, minerals, vitamins, salts, preservatives, antioxidants (such as BHT (butylated hydroxytoluene)), anticaking agents (such as silicon dioxide and maltodextrin), anti-foaming agents and the like, which are suitable for human consumption.

[0064] For example, the dry powder meat protein hydrolysates of the invention, prior to addition of carriers or additives, may comprise a protein content of about 80 wt.%, such as about 85 wt. % or higher, about 87% wt. % or higher, about 88 wt. % or higher, about 89 wt. % or higher or about 90 wt. % or higher. Preferably the hydrolysates of the invention comprise a fat content of about 1 wt.% or lower, about 0.9 wt % or lower, about 0.8 wt % or lower, about 0.7 wt % or lower, about 0.6 wt % or lower or about 0.5 wt % or lower.

[0065] In another aspect, the meat protein hydrolysates of the invention are provided in aqueous solution form, which is a form that is particularly suitable for incorporation into liquid products such as beverages, soups, sauces and the like. Aqueous solution hydrolysates of the invention can be produced by cessation of the production process prior to concentration and drying steps or by rehydrating dry powder hydrolysates of the invention with aqueous solution. The aqueous solution comprises water, such as filtered and / or deionised water, and may include other soluble components such as salts, acid, alkali, buffer, vitamins, minerals, colours, flavours, preservatives or other additives that are suitable for human consumption, depending upon the intended end use of the material.

[0066] In one aspect, the process for production of the meat protein hydrolysates of the invention may comprise process steps as outlined below, noting that the present invention is not limited to hydrolysates obtained by this method. It may be possible to modify the sequence of steps, alter treatment parameters, remove one or more steps and / or add one or more additional steps and still produce a meat protein hydrolysate product that is highly soluble in aqueous solution, exhibits low bitterness and is suitable for human consumption. Standard steps in a process conceived by the present inventors to produce the hydrolysates of the invention comprise:a) Adding water or aqueous solution to ground or minced meat to produce an aqueous meat slurry. The proportion of minced meat to aqueous solution can be varied as desired. For example, the wt.% of meat in the slurry may be from about 5 wt.% to about 50 wt.%, such as from about 10 wt.% to about 45 wt.% , from about 15 wt.% to about 40 wt.% or about 20 wt.%, about 25 wt.%, about 30 wt.%, about 35 wt.%, about 40 wt.% or about 45 wt.%. The aqueous solution can include soluble components such as salts, acid, alkali or buffer, such as sodium hydroxide, ammonium hydroxide, sodium bicarbonate, potassium hydroxide, magnesium hydroxide, phosphoric acid, citric acid, acetic acid, tartaric acid, lactic acid and folic acid, for example.b) Heating the aqueous meat slurry to from about 60 °C to about 75 °C for about 10 mins to about 90 mins to produce a heated aqueous meat slurry. For example this pretreatment heating may be conducted at about 60 °C, about 62 °C, about 65 °C, about 66 °C, about 67 °C, about 68 °C, about 69 °C or about 70 °C, about 71 °C, about 72 °C, about 73 °C or about 75 °C for about 10 mins, about 15 mins, about 20 mins, about 25 mins, about 30 mins, about 35 mins, about 40 mins, about 45 mins, about 50 mins, about 55 mins, about 60 mins, about 70 mins, about 80 mins and about 90 mins. Without wishing to be bound by theory the inventors believe that this pretreatment heating initiates the unravelling of protein secondary and tertiary structures, preferably without causing any significant heat- initiated hydrolysis. It is believed by the inventors that better results will be achieved at higher temperatures in the range if exposure is for a shorter duration and with lowertemperatures in the range for a longer duration. Some examples of suitable pretreatment temperatures and duration combinations include 60 °C for 90 mins, 65 °C for 60 mins, 70 °C for 30 mins and 75 °C for 15 mins.c) Adjusting temperature of the heated aqueous meat slurry to protease functional temperature and adding sufficient protease to effect a first hydrolysis step, with stirring, over a period of from about 1 hr to about 3 hrs, for example. The temperature and treatment duration selected for the first hydrolysis step will depend upon the optimal conditions for the particular protease adopted. For example, the protease may be a broad specificity protease in the form of an endoproteinase, which has the effect of breaking proteins into smaller peptides by cleaving non-terminal peptide bonds. Examples of suitable endoproteinases include neutral endoproteinase such as FoodPro PNL (from Connell Bros Co), microbial endoproteinase such as Protamex (from Novozymes), subtilase such as Alcalase, and Bromelain. Preferably the meat slurry will be adjusted to the optimal operating temperature of the protease prior to protease addition and will then be held at that temperature for from about 5 mins to about 60 mins before addition of the protease. In some embodiments protease treatment is conducted at from about 55 °C to about 65 °C, such as about 56 °C, about 57 °C, about 58 °C, about 59 °C, about 60 °C, about 61 °C, about 62 °C, about 63 °C or about 64 °C for about 60 mins, about 70 mins, about 80 mins, about 90 mins, about 100 mins, about 110 mins or about 120 mins.Optionally, after the first hydrolysis step a protease deactivation step will be performed such as by heating of the product of the first hydrolysis step to a temperature of about 80 °C to about 90 °C for from about 10 mins to about 20 mins, preferably at about 85 °C for about 15 mins.d) Exposing a product of the first hydrolysis step to sufficient peptidase to effect a second hydrolysis step, with stirring, over a period of from about 30 mins to about 2 hrs. In the case where a protease deactivation heating step has been conducted after the first hydrolysis step, it will be necessary to adjust the temperature of the product of the first hydrolysis step to a functional temperature of the peptidase being utilised. For example, the peptidase may be an exopeptidase, which has the effect of breaking peptides into smaller peptides and amino acids by cleaving terminal peptide bonds. Examples of suitable exopeptidases include fungal exopeptidases (aminopeptidase) such as Flavourzyme (from Novozymes) and Fungal Protease II (from Connell Bros Co). In some embodiments peptidase treatment in hydrolysis step 2 is conducted at from about 55 °C to about 65 °C, such as about 56 °C, about 57 °C, about 58 °C, about 59 °C, about 60 °C, about 61 °C, about 62 °C, about 63 °C or about 64 °C for about 30 mins, about 40 mins, about 50 mins, about 60 mins, about 70 mins, about 80 mins, about 90 mins, about 100 mins, about 110 mins or about 120 mins. In one aspect of the invention the first and second hydrolysis steps are conducted simultaneously. However, it is generallyconsidered by the inventors that improved hydrolysis and removal of bitter peptides is achieved when the first and second hydrolysis steps are conducted separately and sequentially, optionally with a protease inactivation step conducted in between.e) Treating a product of the second hydrolysis step to deactivate the protease and / or peptidase to produce a meat hydrolysate mixture. Similar to deactivation of the protease as outlined above, this step can for example be achieved by heating of the product of the second hydrolysis step to a temperature of about 80 °C to about 90 °C for from about 10 mins to about 20 mins, preferably at about 85 °C for about 15 mins.f) Exposing the meat hydrolysate mixture to one or more separation and / or purification steps to substantially remove solids and fats and produce meat protein hydrolysate. Such steps can also concentrate the hydrolysate material to produce a higher concentration solution and / or to produce a hydrolysate in a dry powdered form.

[0067] A range of known separation and purification steps are comprehended, which may include one or more of those mentioned below, noting that the order of steps may be modified and that steps may be removed or added to those specifically recited. Generally, although not essentially, it will be preferable to adopt separation I purification steps so as to remove larger impurities and then progressively to remove smaller impurities. Examples of separation I purification steps that may be adopted include:i. filtration using 100 pm filter socks, for example at about 60 °C - 65 °C for about 1 hour and collecting the filtrate;ii. clarification using a clarifier or a decanter to remove fine particles from the filtrate, for example at 60 °C - 65°C for about 2 hours and collecting the supernatant; iii. passing the supernatant though a fat separator and collecting the skim;iv. exposing the skim to activated carbon treatment and collecting the permeate; v. exposing the permeate to ultrafiltration, for example with at least 10kDa size exclusion membrane and collecting the ultrafiltrate.

[0068] In some embodiments the ultrafiltrate is then exposed to a nanofiltration step. The meat protein hydrolysate produced by the one or more separation and / or purification steps may also be exposed to pasteurisation, for example by heating to 63 °C to 65 °C for about 30 min to about 45 min, heating to about 75 °C to about 78 °C for about 8 min to about 10 min or heating to about 85 °C to about 90 °C for about 5 seconds to about 30 seconds, for example.

[0069] In some aspects of the invention the hydrolysate produced by the one or more separation and / or purification steps, or the pasteurized meat protein hydrolysate, is concentrated to produce a concentrated meat protein hydrolysate solution, for example by evaporation using conventional techniques. If it is desired to produce a dry powdered hydrolysate, the concentrated meat protein hydrolysate solution can be dried (via spray drying or other drying technique / s) to produce a meat protein hydrolysate powder. The dry powdered meat hydrolysate can be milled ifrequired using conventional milling techniques and may be packaged for distribution. The products produced by the process outlined above, whether in aqueous solution or in the form of a dry powder, can for example be incorporated into food, beverage or supplement products for human consumption. For example, products incorporating the meat protein hydrolysates of the invention may take the form of a protein drink; a protein supplement powder; a protein bar; a soup or sauce; a pre-prepared meal or weight gain or weight loss supplement / food.EXAMPLES

[0070] The present invention will now be described with reference to the following examples, which should be considered in all respects as illustrative and non-restrictive.EXAMPLE 1 - Laboratory scale general process for production of meat protein hydrolysate

[0071] The meat protein hydrolysis process of the present invention can be carried out at the lab scale according to the following general process:1. Beef trim (10 kg, 92 CL) (which is also referred to as manufacturing grade meat, which is derived from multiple animal muscles) is acquired from a local abattoir and minced through a 4 mm plate before being mixed by hand. Portions of mince (250 g) are then vacuum packaged and stored at -20°C. Frozen portions of minced beef were defrosted overnight at 4 °C prior to hydrolysis.2. Mince (~20 g) is weighed into 250 mL Schott bottles, in duplicate, before the addition of 80 mL of deionised water to produce a 20 wt.% slurry. NH4OH, 28-30 % is added to bring the pH of the solution to 7.0.3. A pretreatment step is then performed by placing the samples in a 70 °C water bath for 30 min.4. All samples are tempered to 60 °C prior to the addition of a neutral protease (FoodPro PNL, Connell Bros Co) (Enzyme A) at 0.2 wt.% (relative to meat weight).5. Hydrolysis is undertaken for 2 h at 60 °C and mixing is achieved by a submersible magnetic stirrer at a speed of 350 rpm.6. Aminopeptidase (Flavourzyme 1000 L, Novozymes) (Enzyme B) is added (0.15 wt.% (relative to meat weight) at 1 h for all treatments.7. Enzyme deactivation is achieved by placing the samples into a water bath at 85 °C for 15 min prior to rapid cooling of the hydrolysates in an ice bath.8. The cooled hydrolysates are then filtered through a Chux™ kitchen cloth placed on top of a Whatman #4 filter paper into 100 mL storage containers.9. Sample aliquots of hydrolysate are weighed into fared petri dishes and frozen at -80 °C.Samples are then dried in a laboratory freeze drier set to -35 °C.10. Hydrolysate samples are frozen at -20 °C prior to degree of hydrolysis (%), total soluble solids (%) and sensory analysis.Degree of Hydrolysis measurement:

[0072] Degree of hydrolysis (DH %) was assayed as per the method of Nielsen et al. (2001) (Ref. 4). Briefly, 400 pL of diluted hydrolysate was added to a cuvette containing 3 mL of o-phthaldialdehyde (OPA) reagent, mixed, and read at 340 nm in a UV / Vis spectrometer exactly 2 min after the sample addition. All samples were assayed in triplicate. Samples were compared to a standard solution of the amino acid serine, to determine milliequivalents serine NH2 / g protein before the DH % was calculated as the percentage of hydrolysed peptide bonds to total peptide bonds. It is reported that the DH % and bitterness level has a bell curve relationship where hydrolysates with low or high DH % values are considered less bitter (Liu et al 2022) (Ref. 5). A DH % of from about 8 % to about 20 % is generally acceptable and DH % of from about 10 % to about 15 % have generally been shown through the inventors’ work to be favourable from a bitterness perspective in hydrolysates prepared from meat substrate using the lab scale process. DH % values of 20 - 25 % are often (but not always) considered unduly bitter.Total Soluble Solids measurement:

[0073] Total soluble solids (TSS %) was estimated by refractive index using an Atago PAL Rl refractometer from a standard curve generated from hydrolysates with known total soluble solids from previous work. Higher TSS results are indicative of a high yield of soluble hydrolysate compounds from the starting substrate but are dependent on the weight of substrate and hydrolysis water used. While higher yields may be favourable economically, higher TSS values can lead to more bitter end products. In hydrolysates prepared from 20 and 40 wt.% meat slurries the inventors have shown from the sensory trials conducted that TSS % values of up to about 6%, and preferably from about 2.5 to about 3.5%, are generally the most acceptable, particularly from a bitterness perspective.Consumer Sensory (Bitterness) Analysis:

[0074] Sensory analysis was undertaken with a panel of 5 project members over four tasting sessions, with 6 hydrolysates being tasted in each session. Panelists were provided with approximately 3 mL of hydrolysate in 30 mL cups that had been tempered to room temperature and the samples were tasted in random order. Water and plain crackers were provided for palette cleansing between samples. Panelists were asked to rate their perceived levels of beefy flavour, saltiness, and bitterness intensity from 0 to 10 and to note other flavours that were present and overall comments. Perceived bitterness is an undesirable trait in protein hydrolysates affecting the acceptability and overall quality of the final product. In the inventors’ experience, hydrolysates with bitterness ratings of 3 or lower are considered the most acceptable. Bitterness ratings above 5 are generally undesirable.

[0075] Variations of the general process in steps 1-10 above were conducted in Examples 2-26 as follows. The same DH %, TSS % and Bitterness analyses were performed in these examples.EXAMPLE 2 - Modified laboratory scale process for production of meat protein hydrolysate where Enzyme A is replaced with Alcalase (Step 4)

[0076] The same process as the general process in steps 1-10 above was conducted, but with the modification that in Step 4 Enzyme A was replaced with alkaline protease (Alcalase, Novozymes) in the amount of 0.2 % w / w of meat.EXAMPLE 3 - Modified laboratory scale process for production of meat protein hydrolysate where Enzyme A is replaced with Protamex (Step 4)

[0077] The same process as the general process in steps 1-10 above was conducted, but with the modification that in Step 4 Enzyme A was replaced with microbial endoproteinase (Protamex, Novozymes) in the amount of 0.2 % w / w of meat.EXAMPLE 4 - Modified laboratory scale process for production of meat protein hydrolysate where Enzyme A is replaced with Alcalase (Step 4)

[0078] The same process as the general process in steps 1-10 above was conducted, but with the modification that in Step 4 Enzyme A was replaced with alkaline protease (Alcalase, Novozymes) in the amount of 0.2 % w / w of meat.EXAMPLE 5 - Modified laboratory scale process for production of meat protein hydrolysate where Enzyme A is replaced with Bromelain (Step 4)

[0079] The same process as the general process in steps 1-10 above was conducted, but with the modification that in Step 4 Enzyme A was replaced with Bromelain (pineapple derived protease) (Connell Bros Co) in the amount of 0.2 % w / w of meat.EXAMPLE 6 - Modified laboratory scale process for production of meat protein hydrolysate where Enzyme A is replaced with Papain (Step 4)

[0080] The same process as the general process in steps 1-10 above was conducted, but with the modification that in Step 4 Enzyme A was replaced with Papain (papaya fruit derived protease) (Connell Bros Co) in the amount of 0.2 % w / w of meat.EXAMPLE 7 - Modified laboratory scale process for production of meat protein hydrolysate using kangaroo meat

[0081] The same process as the general process in steps 1-10 above was conducted, but with the modification that in Step 1 where kangaroo meat (offcuts from a variety of muscles) was used instead of beef trim.EXAMPLE 8 - Modified laboratory scale process for production of meat protein hydrolysate using goat meat

[0082] The same process as the general process in steps 1-10 above was conducted, but with the modification that in Step 1 where goat meat (offcuts from a variety of muscles) was used instead of beef trim.EXAMPLE 9 - Modified laboratory scale process for production of meat protein hydrolysate using chicken meat

[0083] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 1 where chicken meat (commercially available chicken mince) was used instead of beef trim and in Step 240 g of chicken mince was used to produce a 40wt.% slurry.EXAMPLE 10 - Modified laboratory scale process for production of meat protein hydrolysate using liver meat

[0084] The same process as the general process in steps 1-10 above was conducted, but with the modification that in Step 1 where beef liver meat was used instead of beef trim.EXAMPLE 11 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry and no pretreatment

[0085] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry and no pretreatment (Step 3) was performed.EXAMPLE 12 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry and 5 min pretreatment

[0086] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry and the pretreatment (Step 3) was performed for 5 mins instead of 30 mins.EXAMPLE 13 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry and 60 min pretreatment

[0087] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 240 g of beef trim mince was used to produce a 40wt.% slurry and the pretreatment (Step 3) was performed for 60 mins instead of 30 mins.EXAMPLE 14 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry and 90 min pretreatment

[0088] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry and the pretreatment (Step 3) was performed for 90 mins instead of 30 mins.EXAMPLE 15 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry and pretreatment at 40°C

[0089] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry and the pretreatment (Step 3) was performed at 40°C instead of 70°C.EXAMPLE 16 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry and pretreatment at 60°C

[0090] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry and the pretreatment (Step 3) was performed at 60°C instead of 70°C.EXAMPLE 17 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry and pretreatment at 80°C

[0091] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry and the pretreatment (Step 3) was performed at 80°C instead of 70°C.EXAMPLE 18 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry and simultaneous addition of Enzymes A and B

[0092] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry and Enzymes A and B were added simultaneously (Steps 4-6) at 60°C with the pH adjusted to 7, instead of enzymes being added seguentially.EXAMPLE 19 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry, no pretreatment step and simultaneous addition of Enzymes A and B

[0093] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry, no pretreatment heating step (Step 3) was conducted and Enzymes A and B were added simultaneously (Steps 4-6) at 60°C with the pH adjusted to 7, instead of enzymes being added seguentially.EXAMPLE 20 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry and replacement of Enzyme B with Fungal Protease II

[0094] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry, and Enzymes B was replaced with aminopeptidase (Fungal Protease II, Connell Bros Co) in Step 6.EXAMPLE 21 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry, pretreatment for 5 mins and simultaneous addition of Enzymes A and B

[0095] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry, the pretreatment heating step (Step 3) was conducted for 5 mins instead of 30 mins and Enzymes A and B were added simultaneously (Steps 4-6) at 60°C with the pH adjusted to 7, instead of enzymes being added sequentially.EXAMPLE 22 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry, pretreatment for 60 mins and simultaneous addition of Enzymes A and B

[0096] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry, the pretreatment heating step (Step 3) was conducted for 60 mins instead of 30 mins and Enzymes A and B were added simultaneously (Steps 4-6) at 60°C with the pH adjusted to 7, instead of enzymes being added sequentially.EXAMPLE 23 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry, pretreatment for 90 mins and simultaneous addition of Enzymes A and B

[0097] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry, the pretreatment heating step (Step 3) was conducted for 90 mins instead of 30 mins and Enzymes A and B were added simultaneously (Steps 4-6) at 60°C with the pH adjusted to 7, instead of enzymes being added sequentially.EXAMPLE 24 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry, pretreatment at 40°C and simultaneous addition of Enzymes A and B

[0098] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry, the pretreatment heating step (Step 3) was conducted at 40°C instead of 70°C and Enzymes A and B were added simultaneously (Steps 4-6) at 60°C with the pH adjusted to 7, instead of enzymes being added sequentially.EXAMPLE 25 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry, pretreatment at 60°C and simultaneous addition of Enzymes A and B

[0099] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry, the pretreatment heating step (Step 3) was conducted at 60°C instead of 70°C and Enzymes A and B were added simultaneously (Steps 4-6) at 60°C with the pH adjusted to 7, instead of enzymes being added sequentially.EXAMPLE 26 - Modified laboratory scale process for production of meat protein hydrolysate using 40wt.% meat slurry, pretreatment at 80°C and simultaneous addition of Enzymes A and B[000100] The same process as the general process in steps 1-10 above was conducted, but with the modifications that in Step 2, 40 g of beef trim mince was used to produce a 40wt.% slurry, the pretreatment heating step (Step 3) was conducted at 80°C instead of 70°C and Enzymes A and B were added simultaneously (Steps 4-6) at 60°C with the pH adjusted to 7, instead of enzymes being added sequentially.EXAMPLE 27 - Modified laboratory scale process for production of meat protein hydrolysate with no pretreatment step and simultaneous addition of Enzymes A and B[000101] The same process as the general process in steps 1-10 above was conducted, but with the modifications that no pretreatment heating step (Step 3) was conducted and Enzymes A and B were added simultaneously (Steps 4-6) at 60°C with the pH adjusted to 7, instead of enzymes being added sequentially.Results from Examples 1-27[000102] The degree of hydrolysis (DH %), total soluble solids (TSS %) and bitterness rating (Bitterness) determined for the meat hydrolysate products from each of Examples 1-26 are shown in Table 1 below:Table 1[000103] In Table 1 the * symbol in the Bitterness column denotes that the meat protein hydrolysate exhibits a bitterness score of above 5, which is associated with extreme bitterness. Bitterness scores of less than about 3 are most desirable.EXAMPLE 28 - Pilot plant scale general process for production of meat protein hydrolysate using beef[000104] The meat protein hydrolysis process of the present invention can be carried out at the pilot plant scale according to the following general process:1. 60 kg of raw beef meat (minced) is obtained and refrigerated at < 5°C for 1-2 days prior to further processing.2. The meat is mixed with 240 kg of water (20wt.%) to make a slurry.3. NaOH is added to the slurry until a pH of 7.0 is achieved.4. The slurry is heated to 70°C for 30 minutes.5. The slurry is tempered to 60°C before the addition of Enzyme A (neutral protease (FoodPro PNL, Connell Bros Co) at 0.2% w / w of meat).6. After 1 hour Enzyme B is added to the mixture (aminopeptidase, Flavourzyme 500 MG, Novozymes at 0.3% w / w of meat). The mixture is continually mixed for a further 1 hour at60°C.7. The slurry is then heated to 85°C for 15 minutes to inactivate the enzymes before being cooled to 60-65°C.8. The slurry is filtered using 100 pm filter socks at 60-65°C for 1 hour. The supernatant (liquid) and the solids are collected separately.9. Clarification is then carried out using KNA3 (GEA Westfalia) separator (clarifier) to remove fine particles at 60-65°C for 2 hours. The supernatant (liquid) and solids are collected separately.10. The supernatant is cooled to 5°C by running through multiple heat exchangers and stored in a refrigerator at < 5°C.11. The stored supernatant is heated to 60-65°C.12. Separation is carried out using a GEA Westfalia separator at 60-65°C for 2 hours.The skim (non-fat liquid) is collected and then the mixture is cooled to 5°C using inline multiple heat exchangers. The cream (fat) is then collected separately.13. The skim (non-fat liquid) is then recirculated at 20-25°C for 1 hour in the flow- through filtration system (Sunmicron HEXB45) with food grade 8wt.% activated carbon pellets (Clarence Water Filters Pty Ltd).14. The permeate is then cooled and stored at < 5°C.15. The mixture is heated to 10°C and then subjected to ultrafiltration (Abcor ultrafiltration plant (10 kDa and 30 kDa size membrane) at 10-20°C is conducted.16. The permeate is pasteurised to 72°C for 15 seconds using plate heat exchanger. 17. The permeate is then cooled to < 5°C.18. The permeate is concentrated to 24% solids using a falling film evaporator (built by CSIRO engineers).19. The concentrated permeate is spray dried (Niro Production Minor Spray dryer). 20. The spray dried powder is then vacuum packed in plastic bags and stored in refrigerated conditions.Yield Measurement:[000105] Yield was determined for the powder by weighing the total powder produced at the end of the process and comparing it to the weight of input material, as follows:Total Powder Out (kg) I Raw Material In (kg) x 100 = Yield %[000106] The yield result and bitterness sensory assessment result (obtained according to the method outlined in Example 1) for the meat protein hydrolysate produced according to trials A, B and C of the pilot scale process are shown in Table 2, below:Table 2 - Pilot scale yield and bitterness results[000107] MDM is derived from meat left on the bone after the de-boning process. This meat is collected by pressing the bone with meat and high connective tissue meat through a machine with holes in it and the soft tissue is squeezed through these holes - resulting in a paste / fine mince like product.EXAMPLE 29 - Pilot plant scale general process for production of meat protein hydrolysate using chicken meat[000108] The meat protein hydrolysis process of the present invention can be carried out at the pilot plant scale according to the following general process:1. 90 kg of raw chicken meat (minced) is obtained and refrigerated at < 5°C for 1-2 days prior to further processing.2. The meat is mixed with 210 kg of water (30wt.%) to make a slurry.3. NaOH is added to the slurry until a pH of 7.0 is achieved.4. The slurry is heated to 70°C for 30 minutes.5. The slurry is tempered to 60°C before the addition of Enzyme A (neutral protease (FoodPro PNL, Connell Bros Co) at 0.2% w / w of meat).6. After 1 hour Enzyme B is added to the mixture (aminopeptidase, Flavourzyme 1000 L, Novozymes at 0.15% w / w of meat). The mixture is continually mixed for a further 1 hourat60°C.7. The slurry is then heated to 85°C for 15 minutes to inactivate the enzymes before being cooled to 60-65°C.8. The slurry is filtered using 100 pm filter socks at 60-65°C for 1 hour. The supernatant (liquid) and the solids are collected separately.9. Clarification is then carried out using KNA3 (GEA Westfalia) separator (clarifier) to remove fine particles at 60-65°C for 2 hours. The supernatant (liquid) and solids are collected separately.10. The supernatant is cooled to 5°C by running through multiple heat exchangers and stored in a refrigerator at < 5°C.11. The stored supernatant is heated to 60-65°C.12. Separation is carried out using a GEA Westfalia separator at 60-65°C for 2 hours.The skim (non-fat liquid) is collected and then the mixture is cooled to 5°C using inline multiple heat exchangers. The cream (fat) is then collected separately.13. The skim (non-fat liguid) is then recirculated at 20-25°C for 1 hour in the flow- through filtration system (Sunmicron HEXB45) with food grade 8wt.% activated carbon pellets (Clarence Water Filters Pty Ltd).14. The permeate is then cooled and stored at < 5°C.15. The mixture is heated to 10°C and then subjected to ultrafiltration (Abcor ultrafiltration plant (10 kDa and 30 kDa size membrane) at 10-20°C is conducted. 16. The permeate is pasteurised to 72°C for 15 seconds using plate heat exchanger. 17. The permeate is then cooled to < 5°C.18. The permeate is concentrated to 24% solids using a falling film evaporator (built by CSIRO engineers).19. The concentrated permeate is spray dried (Niro Production Minor Spray dryer). 20. The spray dried powder is then vacuum packed in plastic bags and stored in refrigerated conditions.EXAMPLE 30 - Size exclusion chromatography analysis of meat protein hydrolysate peptide molecular weight[000109] Meat protein hydrolysate produced according to the laboratory scale general process of Example 1 was subjected to size exclusion chromatography (SEC) using an Agilent AdvanceBio SEC (7.8 x 300 mm, 2.7 pm) column to obtain information regarding the approximate molecular weight range for the pool of peptides present. The SEC chromatography column was calibrated with the molecular weight standard by comparison of elution time against the calibration curve shown in Fig 1 to derive the molecular weight against retention time correlation shown in Tables 3 and 4 below.Table 3 - Molecular weight versus retention time derived from calibration curve (beef powder)Table 4 - Molecular weight versus retention time derived from calibration curve (chicken powder)[000110] After calibration of the SEC column, meat protein hydrolysate product was subjected to chromatographic analysis to determine the approximate molecular weight ranges of peptides present in the hydrolysate. Fig 2 shows the chromatogram obtained from meat (beef) and Tables 5 and 6 below provide quantitative information on relative percentages of product with respect to molecular weight range for both beef and chicken powder.Table 5 - Meat hydrolysate peptide molecular weight analysis - Beef Powder><Table 6 - Meat hydrolysate peptide molecular weight analysis - Chicken Powder><[000111] From the data in Tables 5 and 6, it is apparent that the hydrolysate sample is primarily composed of peptide material of less than 10kDa in molecular weight and in fact has >99% peak area with molecular weight of less than 5 kDa. This data highlights that the hydrolysis process of the invention results in extensive hydrolysis of the meat proteins.EXAMPLE 31 - Solubility Data[000112] All solubilities are greater than or equal to 95 % soluble at pH 2, pH 8 and with no pH adjustment in 5 % solutions of powder.EXAMPLE 32 - Analysis of meat protein hydrolysate produced by pilot scale process of Examples 28 and 29[000113] Assays of the meat protein hydrolysate produced by the pilot scale processes of Examples 28 and 29 were undertaken by the following well-known methods:• Protein - Kjeldahl nitrogen (Protein = total Kjeldahl nitrogen x 6.25)• Total Fat - Mojonnier method• Ash - Gravimetric determination• Minerals - ICPAES / MS• Amino acids - LC-PDA-MS / MS[000114] References for methods used:Amino Acids - LIPLC Amino Acid Analysis Solution System Guide 71500129702 I Revision BElements - USEPA 6010, 6020 & AOAC, 16th Edition, Methods 986.15 and 974.14 Moisture - AOAC 16th Ed. 934.06, 964.22 &AS2300.1.1Protein - AOAC 18th Ed. 981.10, 920.152, 990.03, 920.87 &AS2300.1.2.1 Ash - AOAC 18th Ed. 2005, 923.03 and 900.02Total Fat - AOAC 18th Edition 954.02,948.15,922.08Table 7 - Characteristics of hydrolysate produced according to Example 28 - using beef<Table 8 - Characteristics of hydrolysate produced according to Example 29 - using chicken meat<[000115] Although the invention has been described with reference to specific examples, it will be appreciated by those skilled in the art that the invention may be embodied in many other forms, and in particular features of any one of the various described examples may be provided in any combination in any of the other described examples. Various modifications and alterations to this invention will become apparent to those skilled in the art without departing from the scope and spirit of this invention. It should be understood that this invention is not intended to be unduly limited by the illustrative embodiments and examples set forth herein and that such examples and embodiments are presented by way of example only with the scope of the invention intended to be limited only by the claims set forth herein as follows.REFERENCES1. Sun X, Zheng J, Liu B, Huang Z and Chen F (2022) Characteristics of the enzyme- induced release of bitter peptides from wheat gluten hydrolysates. Front. Nutr.9:1022257. doi: 10.3389 / fnut.2022.10222572. R. E. Aluko (2017). Structural characteristics of food protein derived bitter peptides. In Bitterness: Perception, Chemistry and Food Processing, Editors: Michel Aliani & Michael N. A. Eskin John. Wiley & Sons, Inc. pp 105-1293. Jens Adler-Nissen (1986). Enzymic hydrolysis of food proteins. Elsevier Sciences.Pages 451.4. Nielsen, P. M., Petersen, D., & Dambmann, C. (2001). Improved method for determining food protein degree of hydrolysis. Journal of Food Science, 66(5), 642-646.5. Liu, B., Li, N., Chen, F., Zhang, J., Sun, X., Xu, L., & Fang, F. (2022). Review on the release mechanism and debittering technology of bitter peptides from protein hydrolysates. Comprehensive Reviews in Food Science and Food Safety, 21(6), 5153- 51706. Anema, S. G., Pinder, D. N., Hunter, R. J., & Hemar, Y. (2006). Effects of storage temperature on the solubility of milk protein concentrate (MPC85). Food Hydrocolloids, 20(2-3), 386-393.)

Claims

CLAIMS1. A meat protein hydrolysate derived from animal meat, wherein the protein hydrolysate is highly soluble in aqueous solution, exhibits low bitterness and is suitable for human consumption.

2. The meat protein hydrolysate according to claim 1, having a degree of hydrolysis (DH) of from about 10% to about 20%.

3. The meat protein hydrolysate according to claim 1 or claim 2, having total soluble solids (TSS) from about 2% to about 6% w / w.

4. The meat protein hydrolysate according to any one of claims 1 to 3, having a bitterness score of about 3 or less.

5. The meat protein hydrolysate according to any one of claims 1 to 4, wherein peptides within the hydrolysate have molecular weight of about 10 kDa or less.

6. The meat protein hydrolysate according to any one of claims 1 to 5, which is in dry powder form.

7. The meat protein hydrolysate according to claim 6, having protein content of about 80% (w / w) or higher and fat content of about 1% (w / w) or lower.

8. The meat protein hydrolysate according to claim 6 or claim 7, which exhibits solubility of 95 % w / w or greater in aqueous solution at pH of from about 2 to about 8.

9. The meat protein hydrolysate according to any one of claims 1 to 5, which is in aqueous solution form.

10. The meat protein hydrolysate according to any one of claims 1 to 9, produced from low- grade meat cuts and / or meat co- or by-products.

11. The meat protein hydrolysate according to any one of claims 1 to 10, wherein the animal meat is bovine, ovine, porcine, poultry, caprine, cameline, macropod or piscine.

12. The meat protein hydrolysate according to any one of claims 1 to 11, which is substantially devoid of egg, milk, nut, grain and legume derived human allergens.

13. The meat protein hydrolysate according to any one of claims 1 to 12, comprising one or more carriers and / or additives.

14. A process for production of a meat protein hydrolysate derived from animal meat, wherein the protein hydrolysate is highly soluble in aqueous solution, exhibits low bitterness and is suitable for human consumption, wherein the process comprises:a) adding water or aqueous solution to ground or minced meat to produce an aqueous meat slurry;b) heating the aqueous meat slurry to from about 60 °C to about 75 °C for from about 10 mins to about 90 mins to produce a heated aqueous meat slurry; c) adjusting temperature of the heated aqueous meat slurry to protease functional temperature and adding sufficient protease to effect a first hydrolysis step, with stirring, over a period of from about 1 hr to about 3 hrs;d) exposing a product of the first hydrolysis step to sufficient peptidase to effect a second hydrolysis step, with stirring, over a period of from about 30 mins to about 2 hrs;e) treating a product of the second hydrolysis step to deactivate the protease and / or peptidase to produce a meat hydrolysate mixture;f) exposing the meat hydrolysate mixture to one or more separation and / or purification steps to substantially remove solids and fats and produce meat protein hydrolysate.

15. The process according to claim 14 wherein steps c) and d) are conducted simultaneously.

16. The process according to paragraph 14 wherein steps c) and d) are conducted separately.

17. The process according to any one of claims 14 to 16 wherein the protease comprises a neutral endoproteinase.

18. The process according to any one of claims 14 to 17 wherein the peptidase comprises an exopeptidase.

19. The process according to any one of claims 14 to 18 wherein in step b) the aqueous meat slurry is heated to about 70 °C for about 30 mins.

20. The process according to any one of claims 14 to 19 wherein the deactivation of the protease and / or peptidase in step e) is achieved by heating the product of the second hydrolysis step to from about 80 °C to about 90 °C for from about 10 mins to about 20 mins.

21. The process according to any one of claims 14 to 20 wherein the one or more separation and / or purification steps comprise:i. filtration using 100 pm filter socks at 60-65°C for about 1 hour and collecting the filtrate;ii. clarification using a clarifier to remove fine particles from the filtrate at 60-65°C for about 2 hours and collecting the supernatant;iii. passing the supernatant though a fat separator and collecting the skim; iv. exposing the skim to activated carbon treatment and collecting the permeate; v. exposing the permeate to ultrafiltration with a 10 kDa size exclusion membrane and collecting the ultrafiltrate.

22. The process according to claim 21 further comprising a nanofiltration step.

23. The process according to any one of claims 14 to 22 wherein the meat protein hydrolysate produced by the one or more separation and / or purification steps is pasteurized.

24. The process according to claim 23 wherein the pasteurized product is concentrated to produce a concentrated meat protein hydrolysate solution.

25. The process according to claim 24 wherein the concentrated meat protein hydrolysate solution is spray dried to produce a meat protein hydrolysate powder.

26. A meat protein hydrolysate produced by the process according to any one of claims 14 to 25.

27. A food, beverage or supplement product for human consumption comprising the meat protein hydrolysate according to any one of claims 1 to 13 or 26.

8. The product according to claim 27 which is a protein drink; a protein supplement powder;a protein bar; a soup or sauce; a pre-prepared meal or weight gain or weight loss supplement / food.

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